PXD015496

PXD015496 is an original dataset announced via ProteomeXchange.

Title	Membrane modulation of super-secreting “midiBacillus” expressing the major Staphylococcus aureus antigen – a mass-spectrometry based absolute quantification approach.
Description	Bacillus subtilis is a Gram-positive bacterium considered as a “cell-factory” for industrial enzymes and biopharmaceuticals. Given the commercial advantage of this organism, researchers have been working towards the improvement of this organism as a producer of secreted proteins. Nonetheless, there is the need to obtain more knowledge regarding the absolute composition of the gram positive cell membrane in order to increase the secretion capability of B. subtilis and gain insight into potential bottlenecks in protein production and/or secretion. However, membrane proteins are one of the most challenging classes of proteins, mainly due to their high hydrophobicity and low abundance compared to their soluble counterparts. To this regard, we established a method for global characterization of absolute protein abundances within the membrane of B. subtilis and verified the effects of large scale protein export on a super-secreting strain. We did shotgun proteomics on three different fractions (extracellular, cytosol and membrane) and spiked in UPS2 proteins in each of these to calculate absolute protein abundances. The absolute values obtained from the shotgun experiment were validated by targeted proteomics. Furthermore, we performed western blot analysis on the three subcellular fractions using antibodies against proteins localized in these three different compartments in order to visually validate the enrichment of membrane proteins in the corresponding fraction.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_05:01:05.487.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Minia Antelo
SpeciesList	scientific name: Bacillus subtilis subsp. subtilis; NCBI TaxID: 135461;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	TSQ Vantage; LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2019-09-18 05:04:27	ID requested
1	2020-03-20 00:35:21	announced
⏵ 2	2024-10-22 05:01:06	announced	2024-10-22: Updated project metadata.

10.3389/fbioe.2020.00143;

Antelo-Varela M, Aguilar Su, á, rez R, Bartel J, Bernal-Cabas M, Stobernack T, Sura T, van Dijl JM, Maa, ß S, Becher D, Antigen - A Mass-Spectrometry-Based Absolute Quantification Approach. Front Bioeng Biotechnol, 8():143(2020) [pubmed]

curator keyword: Biological

submitter keyword: SRM, Membrane Proteins, Heterologous Protein Production,Proteomics, Absolute Quantification

Dörte Becher
contact affiliation	Institut für Mikrobiologie Universität Greifswald Felix Hausdorff Str.8 17489 Greifswald Tel.: (+49) 03834 420 5903 Fax.: (+49) 03834 420 5002
contact email	dbecher@uni-greifswald.de
lab head
Minia Antelo
contact affiliation	Institute of Microbiology University of Greifswald
contact email	minia.antelovarela@uni-greifswald.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/03/PXD015496

PRIDE project URI

• PRIDE
  1. PXD015496
     1. Label: PRIDE project
     2. Name: Membrane modulation of super-secreting “midiBacillus” expressing the major Staphylococcus aureus antigen – a mass-spectrometry based absolute quantification approach.